LOCUS       sequence-310073-        4571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector with blasticidin-S resistance into ROSA26 targetting 
            vectors .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4571)
  AUTHORS   Schwarz SD, Grundbacher E, Hrovat AM, Xu J, Kusnierczyk A, Vagbo CB,
            Schar P, Schuermann D
  TITLE     Inducible TDG knockout models to study epigenetic regulation.
  JOURNAL   F1000Res. 2020 Sep 9;9:1112. doi: 10.12688/f1000research.25637.2. 
            eCollection 2020.
  PUBMED    33082936
REFERENCE   2  (bases 1 to 4571)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "F1000Res.";
            date: "2020-09-9"; volume: "9:1112. doi"; pages: " 
            10.12688/f1000research.25637.2. eCollection 2020"
COMMENT     Sequence Label: pBigTB
FEATURES             Location/Qualifiers
     source          1..4571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..330
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      181..316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     243..262
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        378..425
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             444..842
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    1000..1121
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1037..1056)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1091..1110
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1138..1154)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1139..1155)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1192..1209)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1198..1422
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(1494..1512)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1533..1549)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1533..1549)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1546..1568)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1557..1573
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1581..1587,1588..1605,1606..1611))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1626..1647
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1764..1781)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1935..2523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2015..2034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2694..3485,3486..3554))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3317..3336
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3555..3659)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3686..4141
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3773..3792)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3983..4004
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4267..4289
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4281..4298
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4282..4298
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4305..4324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4305..4323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(4351..4416,4417..4454)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     primer_bind     complement(join(4567..4571,1..16))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
       61 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      241 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
      301 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
      361 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
      421 acgacaaggt gaggaactaa accatggcca agcctttgtc tcaagaagaa tccaccctca
      481 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
      541 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
      601 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
      661 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
      721 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
      781 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
      841 aagcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
      901 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
      961 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     1021 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1081 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg ctagcatcga
     1141 taccgtcgac ctcgaggggc ccccgcgggc ggccgcgagc tcgctgatca gcctcgactg
     1201 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1261 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1321 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1381 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     1441 ccagctgggg ctcgatcctc tagttggcgc gccaacggta ccagcttttg ttccctttag
     1501 tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1561 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1621 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1681 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1741 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1801 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1861 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1921 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1981 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2041 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2101 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2161 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2221 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2281 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2341 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2401 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2461 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2521 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2581 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2641 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2701 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2761 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2821 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2881 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2941 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3001 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3061 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3121 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3181 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3241 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3301 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3361 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3421 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3481 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3541 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3601 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3661 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3721 gtggtggtta cgcgcagcgt gactgctaca cttgccagcg ccctagcgcc cgctcctttc
     3781 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3841 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3901 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3961 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4021 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4081 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     4141 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4201 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     4261 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat
     4321 agggcgaatt ggccttaatt aagggatctg tagggcgcag tagtccaggg tttccttgat
     4381 gatgtcatac ttatcctgtc cctttttttt ccacagctcg cggttgagga caaactcttc
     4441 gcggtctttc cagtggggat cgacggtatc gtagagtcga ggccgctcta gaactagttg
     4501 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     4561 ttctgtggaa t
//
